Shared Resource Cores

项目SUMMARY（参见说明）： 糖缀合物结构/功能的分析需要来自不同学科的专业知识。 这些工具和方法中的许多不仅困难，而且需要昂贵和复杂的设备。我们已经建立了我们的共享资源核心，不仅包含对本PEG目标最重要的组成部分，而且还建立在我们参与研究者的独特优势之上。每个子核心在其单独的部分中详细说明。 子核心01 -质谱-货车Eyk和Zhang博士是蛋白质和糖蛋白质谱分析的领导者。内核的装备非常精良核心的产品包括：1）蛋白质鉴定; 2）使用糖肽固相萃取对N-连接或O-连接糖蛋白进行整体定量; 3）凝集素亲和力/MS分析; 4）通过凝集素微阵列监测糖基化变化; 5）使用“点击”化学分析代谢产生的糖缀合物; 6）通过MALDI-MS”（AXIMA-共振）和聚糖分析仪进行聚糖分析。7)用于分离糖蛋白的细胞表面捕获技术; 2）从细胞质或细胞外环境中靶向捕获可溶性糖蛋白; 3）用于靶向定量糖蛋白和糖基化位点的MRM MS/MS分析。 子核心C2 - NMR -如果可能，聚糖结构最好通过NMR确定。艾伦布什博士是通过核磁共振进行聚糖结构分析的领先专家。他的子核心通过提供寡糖、多糖、糖肽、小糖蛋白和糖脂的结构专业知识来支持PEG。在该次堆芯中可使用最先进的核磁共振设备和方法。 亚核心C3 -化学-Kevin Yarema博士是一位领先的化学糖生物学家。他的子核心将通过以下方式支持PEG：1）为“代谢糖工程”提供糖类似物（例如，SCFA-己糖胺）; 2)进行抑制剂的定制合成（例如，D-PdR、TMG）、叠氮糖和糖核苷酸、聚糖标记工具和其他小分子。 子核心04 - 0-GlcNAc资源-哈特博士和Zachara博士是0-GlcNAc分析的领导者。 为了支持PEG，我们提供：1）检测0-GlcNAc、OGT和OGA的工具; 2）开发位点特异性0-GlcNAc抗体; 3）提供合成0-GlcNAc肽用于抗体生产和MS标准品; 4）OGT和OGA活性测定; 5）糖核苷酸和核苷酸定量; 6）协助单个蛋白或全球的位点定位和定量。这一共享资源提供了关键技术，使糖科学研究的快速发展，对NHLBI的使命至关重要。

PROJECT SU MMARY (See instructions): Analysis of glycoconjugate structure/functions requires specialized expertise from various disciplines. Many of these tools and approaches are not only difficult, but also require expensive and complex equipment. We have established our Shared Resource Core to not only contain the components that are most essential for the aims of this PEG, but also that build upon the unique strengths of our participating investigators. Each sub-Core is detailed in their separate sections. Sub-Core 01 - Mass Spectrometry -Drs. van Eyk and Zhang are leaders in mass spectrometric analysis of proteins and glycoproteins. The Core is extraordinarily well equipped. The Core's offerings include: 1) Identification of proteins; 2) global quantification of N-linked or O-linked glycoproteins using solidphase extraction of glycopeptides; 3) Lectin affinity/MS analyses; 4) Monitoring changes in glycosylation by lectin microarrays; 5) The use of "Click" chemistry for analyses of metabolically produced glycoconjugates; 6)Glycan profiling by MALDI-MS" (AXIMA-Resonance) and GlycanAnalyser. 7) Cell surface capture technology for isolation of glycoproteins; 2) Targeted capture of soluble glycoproteins from cytoplasm or extracellular environment; 3) MRM MS/MS assays for targeted quantification ofglycoproteins and glycosylation sites. Sub-Core C2 - NMR - When possible, glycan structure is best determined by NMR. Dr. Allen Bush is a leading expert in the structural analysis of glycans by NMR. His sub-Core supports the PEG by offering structural expertise for oligosaccharides, polysaccharides, glycopeptides, small glycoproteins and glycolipids. State of the art NMR equipment and methods are available in this sub-Core. Sub-Core C3 - Chemistry - Dr. Kevin Yarema is a leading chemical glycobiologist. His sub-Core will support the PEGs by: 1) providing sugar analogs for "metabolic glycoengineering" (eg. SCFA-hexosamines); 2) performing custom synthesis of inhibitors (eg. D-PDMP, TMG), azido sugars and sugar nucleotides, glycan-tagging tools and other small molecules. Sub-Core 04 - 0-GlcNAc Resources - Drs. Hart and Zachara are leaders in the analysis of 0-GlcNAc. In support of the PEGs we offer: 1) Tools for detecting 0-GlcNAc, OGT and OGA; 2) Develop site-specific 0-GlcNAc antibodies; 3) Provide synthetic 0-GlcNAc peptides for antibody production and MS standards; 4) Assays for OGT and OGA activity; 5) Quantitation of sugar nucleotides & nucleotides; 6) Assist with site mapping and quantification on single proteins or globally. This Shared Resource provides key technologies to allow rapid advancement of glycoscience research critical to the mission of the NHLBI.